- phenixbb - phenix-online.org

Is DEN refinement working?
by Wallen, Jamie 28 Jun '13

28 Jun '13

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PHASER-TFZ scores with ensembles higher?
by Jan Gebauer 28 Jun '13

28 Jun '13

Dear Phaser users, I recently tried to solve two difficult MR jobs by using ensembles in PHASER. With ENSEMBLES in this case I mean aligned structures prepared for PHASER by Phenix.ENSEMBLER. In both case I got solution with rather high TFZ-score (> 8), which I couldn't get wit individual structure, but was not able to refine them. My question is, whether using ensembles would increase the Z-Score without really improving phases. In other words, if there is another cut-off for the "Phaser solved it" Z-score, when using ensemble in contrast to single structures... Regards, Jan -- Dr. Jan Gebauer AG Prof. Baumann Institut für Biochemie / Uni-Köln Otto-Fischer-Str. 12-14 / 50674 Köln Fon: +49 (221) 470 3212 Fax: +49 (221) 470 5066 http://px.uni-koeln.de/

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refining the anomalous Se in MSE using Phenix Gui
by Kenneth A. Satyshur 27 Jun '13

27 Jun '13

Sirs: I am trying to refine the anomalous data in phenix refine. I am using the GUI and I get a box for the anomalous groups. But I when I put in 'Se' and refine: phenix says 'Sorry'. If I use View, I get all atoms in a viewer. I have 5 molecules of 330 residues each and 25 total Se atoms. I cannot possibly pick them out from the molecular display. Then I tried to make a heavy atom file with just selenium in it, but on input, it is read as a 'input model' and when I refine, it says 'duplicate atoms'. Without the anomalous refine, there is a lot of residual Fo-Fc density around the seleniums in the maps. How do I enter the 'anomalous group' into phenix refine. What is the syntax? I can find no help file or documentation on this. Anyone done this before? thanks -- Kenneth A. Satyshur, M.S.,Ph.D. Senior Scientist University of Wisconsin Madison, Wisconsin 53706 608-215-5207

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Phenix version 1.8.2 released
by Paul Adams 26 Jun '13

26 Jun '13

The Phenix developers are pleased to announce that version 1.8.2 of Phenix is now available. Binary installers for Linux, Mac OSX, and Windows platforms are available at the download site: http://phenix-online.org/download/ Highlights from this version: Ensemble refinement, updated hydrogen parameters in refinement and validation, mmCIF reading and writing, LLG maps output from refinement, ligand pipeline for automated solution of protein/ligand complexes, autobuilding starting from a very accurate but very small part of a model, CC* calculation from unmerged data, maximum entropy maps, and PDB editor. Graphical interface: ==================== - New GUIs: phenix.merging_statistics, phenix.pdb_editor General: ======== - Multiple bug fixes - phenix.refine, phenix.reduce, and MolProbity updated to use consistent, revised, electron-cloud centers for hydrogen positions as default (for X-ray) - also see MolProbity website for details: http://molprobity.biochem.duke.edu - mmCIF format now supported in place of PDB format for input to many programs - new commands: phenix.ensemble_refinement, phenix.ligand_pipeline, phenix.sort_hetatms, phenix.cc_star, phenix.pdb_editor, phenix.maximum_entropy_map Refinement: =========== - phenix.refine - support for LLG map type - uses Phaser to compute residual anomalous LLG map - best when used with group_anomalous strategy, which will flatten the LLG map around existing anomalous scatterers with refined f'/f''. - alternatively, anomalous_residual map type (similar but less sensitive) - automatic linking in refinement and pdbtools for residues in the same chain - enabled with automatic_linking.intra_chain=True; this includes carbohydrate linking to protein and between sugars, and covalently bound ligands. - optional output of mmCIF format model and data files - phenix.ensemble_refinement (new command): - New method for refining ensemble models - Combines MD simulation with X-ray restraints that simultaneously account for anisotopic and anharmonic atomic distributions. - For full description see: Burnley BT, Afonine PV, Adams PD, Gros P. 2012. Modelling dynamics in protein crystal structures by ensemble refinement. eLife 1:e00311. doi: 10.7554/elife.00311. Experimental Phasing and Model Building: ======================================== - phenix.autobuild: - now uses torsion NCS parameterization when NCS is used in refinement - building starting from a very accurate but very small part of a model: You can now use the keyword rebuild_from_fragments=True to start rebuilding from fragments of a model. You might want to use this if you look for ideal helices using Phaser, then rebuild the resulting partial model, as in the Arcimboldo procedure. The special feature of finding helices is that they can be very accurately placed in some cases. This really helps the subsequent rebuilding. If you have enough computer time, then run it several or even many times with different values of i_ran_seed. Each time you'll get a slightly different result. - Base-pairing in RNA building -- phenix.autobuild will now try to guess which bases in a model are base-paired, and if there is no positive sequence match to the model, the bases that are base-paired will be chosen to be complementary. You can set the cutoff for base pairing with the keyword dist_cut_base. - waters are automatically named with the chain of the closest macromolecule if you set sort_hetatms=True. This is for the final model only. - you can supply a target position for your model with map_to_object=my_target.pdb. Then at the very end of the run your molecule will be placed as close to this as possible. The center of mass of the autobuild model will be superimposed on the center of mass of my_target.pdb using space group symmetry, taking any match closer than 15 A within 3 unit cells of the original position. The new file will be overall_best_mapped.pdb. Molecular Replacement: ====================== - phenix.mr_rosetta - You can now give commands for Rosetta in mr_rosetta, including a command to specify where disulfide bonds are located - The Rosetta models are now identified by an ID number so that they have unique names - Default number of homology models to download is now 1 (was 5) - Default number of NCS copies (if ncs_copies=Auto) is number leading to solvent content closest to 50%; if ncs_copies=None then all plausible values of ncs_copies are tested. New commands: ============= - phenix.ligand_pipeline (new command): - combines Xtriage, Phaser, eLBOW, phenix.refine, AutoBuild, and LigandFit to automatically solve protein/ligand complexes - optional integration with Coot (parameter interactive=True) allows semi-interactive operation where more rebuilding is required - phenix.cc_star (new command): - combined assessment of model and data quality, as outlined in Karplus & Diederichs (2012) Science 336:1030-3. - summary also output by phenix.model_vs_data if additional unmerged_data keyword is specified - phenix.maximum_entropy_map (new command): - tool to compute maximum entropy map from map coefficients. The program reads input Fourier map coefficients and modifies corresponding synthesis using Maximum-Entropy Method (MEM). The MEM modified map is everywhere positive, smooth and is of higher resolution. The method uses is a modification of Gull & Daniell (1978) algorithm. - phenix.sort_hetatms (new command): - rearranges heteroatoms (ligands, waters, etc.) in a model so they are paired with the nearest macromolecule chain, similar to the PDB's processing of models - phenix.pdb_editor (new command): - graphical editor for PDB files, based on tree view of model hierarchy - perform common operations such as chain renaming, renumbering, manipulation of atomic properties, add/remove atoms Ligands: ======== - phenix.ready_set: - methyl rotations now automatic when adding explicit hydrogens (also in phenix.reduce) - more options adding deuteriums to a model have been introduced Miscellaneous: ============== - phenix.xtriage: - include merging statistics if unmerged intensities are used as input - phenix.maps: - added wavelength parameter and support for anomalous residual and Phaser LLG map coefficients For a full list of changes see: http://www.phenix-online.org/documentation/CHANGES Please note that this publication should be used to cite use of Phenix: PHENIX: a comprehensive Python-based system for macromolecular structure solution. P. D. Adams, P. V. Afonine, G. Bunkóczi, V. B. Chen, I. W. Davis, N. Echols, J. J. Headd, L.-W. Hung, G. J. Kapral, R. W. Grosse-Kunstleve, A. J. McCoy, N. W. Moriarty, R. Oeffner, R. J. Read, D. C. Richardson, J. S. Richardson, T. C. Terwilliger and P. H. Zwart. Acta Cryst. D66, 213-221 (2010). Full documentation is available here: http://www.phenix-online.org/documentation/ There is a Phenix bulletin board: http://www.phenix-online.org/mailman/listinfo/phenixbb/ Please consult the installer README file or online documentation for installation instructions. Direct questions and problem reports to the bulletin board or: help(a)phenix-online.org and bugs(a)phenix-online.org Commercial users interested in obtaining access to Phenix should visit the Phenix website for information about the Phenix Industrial Consortium. The development of Phenix is principally funded by the National Institute of General Medical Sciences (NIH) under grant P01-GM063210. We also acknowledge the generous support of the members of the Phenix Industrial Consortium. -- Paul Adams Deputy Division Director, Physical Biosciences Division, Lawrence Berkeley Lab Division Deputy for Biosciences, Advanced Light Source, Lawrence Berkeley Lab Adjunct Professor, Department of Bioengineering, U.C. Berkeley Vice President for Technology, the Joint BioEnergy Institute Laboratory Research Manager, ENIGMA Science Focus Area Building 64, Room 248 Tel: 1-510-486-4225, Fax: 1-510-486-5909 http://cci.lbl.gov/paul Lawrence Berkeley Laboratory 1 Cyclotron Road BLDG 64R0121 Berkeley, CA 94720, USA. Executive Assistant: Louise Benvenue [ LBenvenue(a)lbl.gov ][ 1-510-495-2506 ] --

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what about heavy atom as model for MR
by 赵岩 26 Jun '13

26 Jun '13

Hi everyone, The heavy atoms(Se-Met) were located at P4322 space group(4A). But the initial density map is so bad that I can not build the initial model. Meanwhile, I have another two datasets, native(2.6A) and se-met anomalous dataset with space group P2221. But these two datasets have no solution. I have tried the initial map from sp P4322 as a model to do molecular replacement. But no MR solution. So, I want to know how can I use the heavy atom as model to do MR with isomorphous or anomalous differences? Thanks! -- Yan Zhao, M.Phil. National Laboratory of Protein Sciences Institute of Biophysics Chinese Academy of Sciences 15 Datun Rd. Beijing, 100101 China

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possible Xtriage bug??
by Michael Thompson 25 Jun '13

25 Jun '13

Hello Developers, I would like to report a possible bug in newer versions of phenix.xtriage. I have noticed recently that I am getting some strange results for the results of the L-test when I run xtriage, as described below: My data show a strong non-crystallographic translation, described by the vector <uvw>=(1/3, 2/3, 1/2). This NCS translation is readily identified by xtriage. When Xtriage performs the L-test, the output states: L test for acentric data using difference vectors (dh,dk,dl) of the form: (2hp,2kp,2lp) where hp, kp, and lp are random signed integers such that 2 <= |dh| + |dk| + |dl| <= 8 Mean |L| :0.504 (untwinned: 0.500; perfect twin: 0.375) Mean L^2 :0.340 (untwinned: 0.333; perfect twin: 0.200) These results don't make sense, as the statistics show an "anti-twinned" effect, which should not happen with the L-test. It appears that xtriage is using reflections that do not have the same expected intensity values due to the NCS translation. The strange thing is that I have another xtriage logfile, which I produced in September of 2012 using the same input data, that indicates the reflections used for the L-test were chosen "using difference vectors (dh,dk,dl) of the form: (3hp,3kp,2lp)." This choice of reflections does account for proper parity, and I get results that are much more sensible (Mean |L|=0.394; Mean L^2=0.221). I am wondering why I am getting different results now than I did last September? Has something changed in the underlying code, or am I missing something silly? I do not have an easy way of determining which version of phenix I used when the L-test was conducted using sensible reflections (the xtriage logfile doesn't contain this info as far as I can tell), but the version that seems to choose reflections inappropriately is 1.8.2-1334. Thanks a lot for the help, and for phenix being awesome in general! Mike -- Michael C. Thompson Graduate Student Biochemistry & Molecular Biology Division Department of Chemistry & Biochemistry University of California, Los Angeles miket(a)chem.ucla.edu

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How to treat EDTA on a special position
by Jan Gebauer 25 Jun '13

25 Jun '13

Dear all, I recently solved a structure in R32/H (155). Interestingly some crystal contacts are mediated by an Metal/EDTA complex. The metal -presumably a magnesium- sits on a special position (x,0, 1/2) in the middle of a twofold axis with the EDTA molecule coordinating it. (See "top" view in dropbox image below) If I fit the EDTA/Mg complex in the density, the symmetry will of course generate a symmetry mate on the same spot. (see "side" view in dropbox image below). In phenix.refine, both entities are refined, which of course gives wrongs results. My question is, how to best define the Ligand to be compatible with phenix.refine and -later- with pdb submission. Should I just set occupancy to 0.5 for the whole EDTA molecule? - But then I would still get two molecules at this site... Or split it somehow half and let the second half be generated by symmetry? In that case, I would need to define bonds over the asymmetric unit to prevent "dissociation" of my ligand halves, wouldn't I? Or is there a third way? I already searched the phenixBB, but only found some "old" threads discussing vaguely similar problems. Unfortunately, they didn't help me to answer which way is better... (or right...) Regards, Jan DROPBOX IMAGE: https://dl.dropboxusercontent.com/u/29745394/R32%20-%20EDTA.jpg -- Dr. Jan Gebauer AG Prof. Baumann Institut für Biochemie / Uni-Köln Otto-Fischer-Str. 12-14 / 50674 Köln Fon: +49 (221) 470 3212 Fax: +49 (221) 470 5066 http://px.uni-koeln.de/

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completeness in table1
by Alexander Batyuk 24 Jun '13

24 Jun '13

Dear developers, What could be the reason for discrepancy in completeness reported in XSCALE.LP (94.5% - 100.0%) vs from Table1: 89.57% (37.49%)? Best wishes, Alex -- Alex Batyuk The Plueckthun Lab www.bioc.uzh.ch/plueckthun

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Re: [phenixbb] How to export difference patterson map in PHENIX
by Ulrich Baumann 20 Jun '13

20 Jun '13

Hi Nat I tried to limit the resolution by either --diff_limit 4 or --high_resolution 4.0. This does not work - probably is not supposed to work in this way? Thanks, ULrich lrich-Baumanns-iMac:xds_new ubaumann$ cctbx.patterson_map au1_run1_Rfree.mtz --high_resolution 4. !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! !! WARNING - EXPERIMENTAL PROGRAM !! !! !! !! This program is still in development - some functionality may be !! !! missing and/or untested. Use at your own risk! For bug reports, etc. !! !! email bugs(a)phenix-online.org. !! !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! Traceback (most recent call last): File "/usr/local/phenix-dev-1405/build/mac-intel-osx-x86_64/../../cctbx_project/iotbx/command_line/patterson_map.py", line 176, in <module> run(sys.argv[1:]) File "/usr/local/phenix-dev-1405/build/mac-intel-osx-x86_64/../../cctbx_project/iotbx/command_line/patterson_map.py", line 136, in run params = cmdline.work.extract() File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 1690, in extract value = object.extract(parent=result) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 1086, in extract return self._type_from_words()(self.words, master=self) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 320, in from_words value = self._value_from_words(words=words, path=master.full_path()) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 348, in _value_from_words return float_from_words(words=words, path=path) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 274, in float_from_words result = number_from_words(words=words, path=path) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 227, in number_from_words value_string=str_from_words(words), words=words, path=path) File "/usr/local/phenix-dev-1405/cctbx_project/libtbx/phil/__init__.py", line 212, in number_from_value_string path, value_string, words[0].where_str())) RuntimeError: Error interpreting high_resolution="True" as a numeric expression (command line argument, line 1) Ulrich-Baumanns-iMac:xds_new ubaumann$ cctbx.patterson_map au1_run1_Rfree.mtz --diff_limit 4 ---------------------------------------- Prof. Ulrich Baumann, PhD Institute of Biochemistry, University of Cologne Otto-Fischer-Strasse 12-14, D-50674 Cologne, Germany

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X-ray/Neutron joint refinement
by Emily Golden 19 Jun '13

19 Jun '13

Hi All, I have a couple of questions about X-ray/neutron joint refinement in phenix. Is it possible to use the neutron data only for refining hydrogen atom positions and the X-ray data for all other atoms? I used the option to add hydrogen atoms to model and refine H/D at exchangable sites and H elsewhere. This worked for many of the exchangeable hydrogen atoms. However, for some side chains, especially Lysine, the program has placed a hydrogen and deuterium atom (with refined occupancies) in the same position as the atom it is supposed to be bonded to. Does anybody know why this is happening and what I may be able to do about it. Or will I need to manually move each misplaced H/D in Coot? Thank you for your help. Emily

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Jerry Karle
by Robert Sweet 19 Jun '13

19 Jun '13

Jerry Karle died a couple of weeks ago, and one of us here learned about it by noticing his obituary in the NY Times: http://www.nytimes.com/2013/06/15/health/jerome-karle-94-dies-nobelist-for-… He's well known to those of us who started in this field when the solving of a Patterson function for a heavy atom in a structure was the only way we knew to get the structure. With Herb Hauptman, and in close teamwork with his wife Isabella who actually >did< crystallography, he played a major role in working out the sort of Direct Methods that allowed one to solve ordinary small-molecule structures before the days of high-speed computing. As the obituary points out, Jerry and Herb shared the 1985 Nobel Prize in Chemistry for this work. The obit. is worth reading. I suspect you'll see an even better one in Nature soon. My own tiny personal memory of Jerry Karle: When I was a grad student at the Univ. of Wisc. with Larry Dahl I made a pilgrimage to the NRL to visit Jerry and Isabella. I was trying to use direct methods to solve at least one cephalosporin structure, which eventually appeared in my thesis, and there was a feature of DM that I failed to understand. They were very patient with me, but couldn't understand what it was I didn't get. It turns out to have been essentially trivial, but that's the way it goes sometimes. Anyway he always recognized me afterwards, or pretended to. I was delighted on arriving at BNL to discover that his daughter Louise was here. Bob ========================================================================= Robert M. Sweet E-Dress: sweet(a)bnl.gov Group Leader, PXRR: Macromolecular ^ (that's L Crystallography Research Resource at NSLS not 1) http://px.nsls.bnl.gov/ Photon Sciences and Biosciences Dept Office and mail, Bldg 745, a.k.a. LOB-5 Brookhaven Nat'l Lab. Phones: Upton, NY 11973 631 344 3401 (Office) U.S.A. 631 344 2741 (Facsimile) =========================================================================

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Re: [phenixbb] cv convert to mtz file
by Pavel Afonine 17 Jun '13

17 Jun '13

Hi Majid, I had no problems converting cbh1_c10a.cv into MTZ using Phenix "Reflection file editor". To convert he other one (cbh1_fg34.cv) you need to add CRYST1 record similarly to cbh1_c10a.cv. Make sure you use recent version of Phenix.. Pavel On 6/17/13 5:48 AM, Majid Haddad Momeni wrote: > Dear Sir/Madam, > > I need help to be able to convert my .cv file to mtz, I have already tried both ccp4 package and Phenix through "Convert to/modify/extend MTZ" and "Reflection file editor" > However they didn't work! > I do highly appreciated of your assistance, > > Best Regards > Majid Haddad Momeni > PhD Student > Swedish University of Agricultural Sciences, > Uppsala-Sweden

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Beamtime available at NE-CAT (APS)
by Cyndi Salbego 14 Jun '13

14 Jun '13

Synchrotron beam time available to general users at NE-CAT (Sector 24 of APS) during the month of July 24ID-C beamline features: - Undulator beamline fully tunable from 6 to 21 keV (2.1 to 0.6 Angstroms) - PILATUS 6M pixel array detector, allowing continuous fine phi-sliced data acquisition at up to 25 frames per second - MD2 microdiffractometer with triple aperture (70, 30, or 10 micron apertures) - MK3 mini-kappa available on the microdiffractometer allowing reorientation of crystals for better diffraction geometry. - ALS style sample mounting robot compatible with ALS pucks or Unipucks 24ID-E beamline features: - Fixed energy undulator beamline (12662 eV) optimized for Se-SAD data collection - Beam stability optimized for smaller beams (down to 5 micron) - ADSC Q315 CCD detector - MD2 microdiffractometer with triple aperture (50, 20, or 5 micron apertures) - ALS style sample mounting robot compatible with ALS pucks or Unipucks - Remote data collection available For more info on our beamlines and updated schedule availability please see necat.chem.cornell.edu or contact Cyndi Salbego (csalbego(a)anl.gov). To apply for General User Time (Rapid Access for 2013-2 run) please go to: https://beam.aps.anl.gov/pls/apsweb/gup0005.start_page Cyndi Salbego Administrator, NE-CAT 630.252.0689

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Re: [phenixbb] elbow
by Mengbin Chen 13 Jun '13

13 Jun '13

Dear Dr. Dias, I tried the command line in Terminal, but Elbow still regularized the conformation and bonds of the ligand and the result turned out to be the same with that I ran thorough GUI. The thing is that the input pdb file has a conformation the ligand should adopt so that I do not really want energy minimization or geometry optimization to be done on it. Although I set "none" for "geometry optimization" in GUI, it unfortunately changed the ligand conformation. Thank you, Mengbin On Thu, Jun 13, 2013 at 2:52 PM, Mengbin Chen <mengbinc(a)sas.upenn.edu>wrote: > Dear Dr. Dias, > > I tried the command line in Terminal, but Elbow still regularized the > conformation and bonds of the ligand and the result turned out to be the > same with that I ran thorough GUI. The thing is that the input pdb file has > a conformation the ligand should adopt so that I do not really want energy > minimization or geometry optimization to be done on it. Although I set > "none" for "geometry optimization" in GUI, it unfortunately changed the > ligand conformation. > > Thank you, > Mengbin > > > On Thu, Jun 13, 2013 at 2:42 PM, Joao Dias <joao.dias(a)heptares.com> wrote: > >> Dear Mengbin, >> You can open a terminal window and run elbow as a command line. >> >> phenix.elbow --final_geometry input_file.pdb**** >> >> >> Do not forget to initialize the phenix environment, depending on your >> terminal shell, one example is: >> source >> /Applications/PHENIX-dev-1320/Contents/phenix-dev-1320/phenix_env.sh >> >> All the best, >> Joao >> >> Dr Joao Dias**** >> >> Principal Scientist**** >> >> ** ** >> >> Heptares Therapeutics Ltd**** >> >> BioPark, Broadwater Road,**** >> >> Welwyn Garden City,**** >> >> Herts, AL7 3AX**** >> >> UK >> >> From: Mengbin Chen <mengbinc(a)sas.upenn.edu> >> Reply-To: PHENIX user mailing list <phenixbb(a)phenix-online.org> >> Date: Thursday, 13 June 2013 19:32 >> To: PHENIX user mailing list <phenixbb(a)phenix-online.org> >> Subject: [phenixbb] elbow >> >> Dear PHENIX Users, >> >> I have been playing around with Elbow for a while, but I kept having >> problems generating .cif file. For "Chemical file type" and "Template PDB >> file", I used the pdb file of the ligand. Since the coordinates in the pdb >> file corresponds to a conformation desired, I chose "None" for "geometry >> optimization". After running Elbow, the output pdb file was completely >> different than the input one -- even double bonds became single bonds and >> 10-membered ring conformation changed. I have tried Sketcher in CCP4 >> package, but no good results either. Could anyone tell me how to do this >> correctly? Any help is appreciated! >> >> Thank you in advance, >> Mengbin >> >> -- >> Mengbin Chen >> Department of Chemistry >> University of Pennsylvania >> >> [image: www.heptares.com] <http://www.heptares.com><http://www.heptares.com> >> >> >> The information in this e-mail is confidential and may be legally >> privileged. It is intended for the exclusive attention of the addressee >> stated above and should not be copied or disclosed to any other. If you >> have received this transmission in error, please make no use of its >> contents and contact the sender. Contact and registered office address: >> Heptares Therapeutics Limited, BioPark, Broadwater Road, Welwyn Garden >> City, Hertfordshire, AL7 3AX. Company No: 06267989 >> >> > > > -- > Mengbin Chen > Department of Chemistry > University of Pennsylvania > -- Mengbin Chen Department of Chemistry University of Pennsylvania

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elbow
by Mengbin Chen 13 Jun '13

13 Jun '13

Dear PHENIX Users, I have been playing around with Elbow for a while, but I kept having problems generating .cif file. For "Chemical file type" and "Template PDB file", I used the pdb file of the ligand. Since the coordinates in the pdb file corresponds to a conformation desired, I chose "None" for "geometry optimization". After running Elbow, the output pdb file was completely different than the input one -- even double bonds became single bonds and 10-membered ring conformation changed. I have tried Sketcher in CCP4 package, but no good results either. Could anyone tell me how to do this correctly? Any help is appreciated! Thank you in advance, Mengbin -- Mengbin Chen Department of Chemistry University of Pennsylvania

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Discrepancy between R-factors from phenix.refine vs phenix Generate "Table 1"
by Sam Stampfer 12 Jun '13

12 Jun '13

Dear Phenix Community, I am preparing my structure for deposition and ran it through the Generate "Table 1" function in phenix. The inputs I gave it were my .mtz structure factors file, the final pdb coordinate file, and the log file from hkl2000. I did not provide unmerged data. My pdb file (which had been refined using phenix.refine) had values of r_work=0.2342 r_free=0.2603 However, phenix.model_vs_data (from Generate "Table 1") gave me r_work=0.2940 r_free=0.3150 I also got this warning about resolution limits, but I think it should not have affected the R-free calculation to this extent: WARNING: Resolution limits in scaling log for structure A549T are inconsistent with resolution limits in the MTZ file: 50.00 - 3.02 (in logfile) 48.47 - 3.03 (in MTZ file) When I refined my model in phenix, I used the twin law h,-h-k,-l. I read in the documentation that twinning can account for some of this discrepancy, but that the program is supposed to take twinning into account if it will lower the calculated R-work by more than 2%, which it doesn't seem to have done (or there is some other problem with my data). Thanks for your help! -Sam

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den refinement
by Andreas Förster 12 Jun '13

12 Jun '13

Dear all, I'm trying to get DEN refinement going from the command line. The run aborts quickly with the following error in the log file: Sorry: Cartesian simulated annealing should not be turned on for DEN refinement. Annealing settings are controlled in the den parameter scope. The DEN scope in input.def looks like this: den { reference_file = den_ref_Jun.pdb gamma = 0.5 kappa = 0.1 weight = 30 sigma = 0.44 optimize = True opt_gamma_values = 0 0.2 0.4 0.6 0.8 1 opt_weight_values = 3 10 30 100 300 num_cycles = 12 kappa_burn_in_cycles = 2 bulk_solvent_and_scale = True refine_adp = True final_refinement_cycle = False verbose = False annealing_type = *torsion cartesian output_kinemage = False restraint_network { lower_distance_cutoff = 3 upper_distance_cutoff = 15 sequence_separation_low = 0 sequence_separation_limit = 10 exclude_hydrogens = True ndistance_ratio = 1 export_den_pairs = False den_network_file = None } Annealing type is torsion from what I can tell. What am I doing wrong? (Release tag: 1394, Platform: intel-linux-2.6-x86_64 redhat-e6.2) Thanks. Andreas -- Andreas Förster, Research Associate Paul Freemont & Xiaodong Zhang Labs Department of Biochemistry, Imperial College London http://www.msf.bio.ic.ac.uk

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Discrepancy between phenix.refine and phenix.model_vs_data R-factors
by Abhinav Kumar 11 Jun '13

11 Jun '13

I am getting a significant difference between the R-factors written by phenix.refine and phenix.model_vs_data. 1. Phenix.refine phenix.refine "in.pdb" "Fobs.mtz" "refinement.input.xray_data.labels=FP SIGFP" "refinement.input.xray_data.high_resolution=2.50" "fix_rotamers=True" "ordered_solvent=True" "strategy=tls+individual_sites+individual_adp" Final R-work = 0.1829, R-free = 0.2141 2. Phenix.model_vs_data phenix.model_vs_data out.pdb Fobs.mtz f_obs_label="FP" r_free_flags_label="FreeR_flag" high_resolution=2.50 comprehensive=true r_work(re-computed) : 0.1808 r_free(re-computed) : 0.2099 How can I reproduce the phenix.refine R-factors in phenix.model_vs_data? -- Thanks, Abhinav JCSG@SSRL, SLAC (650) 926-2992

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phenix elbow: is similar compound helpful to create restraint for new ligand?
by Rongjin 11 Jun '13

11 Jun '13

Hello Phenix users, I have tetrabutylphosphonium in my crystal. I found a similar compound tetrabutylammonium (PDB residue name TBA) which are found in several PDB entries. Usually I draw the new compound in 2-D format and convert to 3D and then create restrain files. In this case, considering N and P are very similar, will the TBS be helpful for elbow to generate restrains for tetrabutylphosphonium? Thanks Rongjin Guan

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Hi
by Wei Shi 10 Jun '13

10 Jun '13

Hi all, I was trying to solve the structure of a protein in several different datasets using xds and phenix. I could solve the structure from one dataset in space group P4. For another dataset, I could solve the structure using the monomer of the structure I got from the first dataset as search model and solve the structure in space group mC. For the third dataset, in IDXREF.LP, the space group of the highest symmetry is hp: 101.2, 101.3, 58.8, 90, 90, 120. According to Mathiews coefficient, 1 monomer is expected in the asymmetric unit. But I couldn't get the molecular replacement solution using the same method as for the second dataset. I also tried several other search models (eg. deletion of the potential flexible region in the search model) and tried to find the solution in all possible pointgroup. I also tried to process the data in oC (C222), the space group of the second highest symmetry in IDXREF.LP, but, still I could not get right molecular replacement solution. I don't whether this means that there is a big conformational change for the structure in the third dataset or the space group I use is not right. Let me know if any of you would have any comments or suggestions for me. Thank you so much! Best, Wei

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Postdoc Position - Berkeley
by Paul Adams 07 Jun '13

07 Jun '13

Lawrence Berkeley Laboratory Postdoctoral Researcher Job ID: 76029 Division: Physical Biosciences (Joint BioEnergy Institute) Date Opened: 6/5/2013 Summary: A postdoctoral position is available at the Joint BioEnergy Institute in Emeryville to work on proteins involved in plant cell wall synthesis. To learn more about JBEI visit: http://www.jbei.org/ Duties: Implementation of methods for the cloning, protein expression, and purification for glycosyl transferases, typically using the Gateway system for gene manipulation. Transformation of host organisms for protein expression, followed by assay of protein expression, and scale up of expression and sample processing (such as deglycosylation). Purification of proteins for biochemical and structural studies. Expression in multiple systems, including E. coli, yeast and insect cells. Qualifications: a Ph.D. in biochemistry, molecular biology, structural biology, or a related field. Demonstrated experience with protein expression, especially in systems other than E. coli. First-author publications demonstrating prior results in the area of protein expression. Experience with cloning methods and protein purification methods. Strong problem solving skills. Solid interpersonal skills and the ability to work in a team environment are critical. Experience with the practical aspects of structure solution by X-ray crystallography would be helpful but not essential. Note that this is a multi-year position that is renewed annually on the basis on performance and continued funding. Lawrence Berkeley National Laboratory is a world leader in science and engineering research, with 13 Nobel Prize recipients over the past 75 years, and 57 present members of the National Academy of Sciences. LBNL conducts unclassified research across a wide range of scientific disciplines and hosts four national user facilities. AA/EEO employer committed to the development of a safe and diverse workforce. Learn more at http://www.lbl.gov. To apply visit https://lbl.taleo.net/careersection/2/jobsearch.ftl?lang=en and search for the job number 76029 or visit: https://lbl.taleo.net/careersection/2/jobdetail.ftl?lang=en&job=76029 -- Paul Adams Deputy Division Director, Physical Biosciences Division, Lawrence Berkeley Lab Division Deputy for Biosciences, Advanced Light Source, Lawrence Berkeley Lab Adjunct Professor, Department of Bioengineering, U.C. Berkeley Vice President for Technology, the Joint BioEnergy Institute Laboratory Research Manager, ENIGMA Science Focus Area Building 64, Room 248 Tel: 1-510-486-4225, Fax: 1-510-486-5909 http://cci.lbl.gov/paul Lawrence Berkeley Laboratory 1 Cyclotron Road BLDG 64R0121 Berkeley, CA 94720, USA. Executive Assistant: Louise Benvenue [ LBenvenue(a)lbl.gov ][ 1-510-495-2506 ] --

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From Dmitri Svergun and Matthias Wilmanns: BSR-2013: Abstract submission deadline is extended until 20th June!
by Terwilliger, Thomas C 07 Jun '13

07 Jun '13

11th International Conference: Biology and Synchrotron Radiation (BSR), 8-11 September 2013, EMBL Hamburg, Germany The abstract submission deadline for BSR-2013 is extended until Thursday 20th June, 2013! Biologists, physicists, chemists, from students to principal investigators, are invited to this tri-annual meeting with rich traditions. Do not miss the unique opportunity to present your work, meet colleagues, friends and future collaboration partners from all over the world. Check out our preliminary programme to learn more. BSR 2013 offers a tour of DESY’s most impressive research infrastructures and includes a broad social programme to discover the vibrant city of Hamburg. Accompanying persons will enjoy special rates! Extended deadlines: ·Registration and abstract submission: 20 June 2013 ·Registration without abstract: 15 August 2013 Take a look at the updates BSR webpage ( www.bsr2013.org<http://www.bsr2013.org/>) for more information and to register. CONTACT: Diah.Yulianti(a)embl.de<mailto:[email protected]>, or margret.fischer(a)embl-hamburg.de<mailto:[email protected]> Alternatively, contact us on the event facebook page

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Re: [phenixbb] Pseudotranslation issue
by Gino Cingolani 07 Jun '13

07 Jun '13

Dear Fulvio, We just had a similar problem (further complicated by some twinning). Assuming that: 1. You collected enough data and 2. You don't have a crazy number of molecules in the tetragonal asymmetric unit, I would advice to scale your data in P1 and do MR in P1. Then use conventional refinement + model building to bootstrap a better model (most likely your initial model isn't as good). Rosetta may help, too. Then eventually go back to the tetragonal data with the improved search model and see if LLG and packing make sense. Gino **************************************************************** Gino Cingolani, Ph.D. Associate Professor Thomas Jefferson University Dept. of Biochemistry & Molecular Biology 233 South 10th Street - Room 826 Philadelphia PA 19107 Office (215) 503 4573 Lab (215) 503 4595 Fax (215) 923 2117 E-mail: gino.cingolani(a)jefferson.edu Website: http://www.cingolanilab.org **************************************************************** "Nati non foste per viver come bruti, ma per seguir virtute e canoscenza" ("You were not born to live like brutes, but to follow virtue and knowledge") Dante, The Divine Comedy (Inferno, XXVI, vv. 119-120) Fulvio Saccoccia <fulvio.saccoccia(a)uniroma1.it> wrote: >Dear phenixbb, > I have a data set collected at 2.2A resolution that I can integrate >in P4. Aimless, Phaser and Xtriage recognized a pseudotranslation >(34.6% of origin peak) at fractional coordinates 0.500 0.500 0.479 >(ORTH 46.7 46.7 37.4). However labelit.index run with >sublattice_allow=true search did not detect the pseudotranslation >vector. So far, any attempts to phase by molecular replacement was >unsuccesfull and I suspect that it could be due to incorrect assignement >of space group: > >Now, some questions arise: >1) does the pseudo translation exist or do not? >2) if translational NCS is present, could it mimic a P41/P42/P43 space >group? >3) in any case, any advices in order to properly deal with tNCS will be >of great help? > >I thank you in advance >Best wishes > >-- >Fulvio Saccoccia, PhD >Sapienza University of Rome >Dept. of Biochemical Sciences "A. Rossi Fanelli" >Piazzale Aldo Moro, 5 >00185 Rome (Italy) > >_______________________________________________ >phenixbb mailing list >phenixbb(a)phenix-online.org >http://phenix-online.org/mailman/listinfo/phenixbb > The information contained in this transmission contains privileged and confidential information. It is intended only for the use of the person named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. CAUTION: Intended recipients should NOT use email communication for emergent or urgent health care matters.

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Pseudotranslation issue
by Fulvio Saccoccia 07 Jun '13

07 Jun '13

Dear phenixbb, I have a data set collected at 2.2A resolution that I can integrate in P4. Aimless, Phaser and Xtriage recognized a pseudotranslation (34.6% of origin peak) at fractional coordinates 0.500 0.500 0.479 (ORTH 46.7 46.7 37.4). However labelit.index run with sublattice_allow=true search did not detect the pseudotranslation vector. So far, any attempts to phase by molecular replacement was unsuccesfull and I suspect that it could be due to incorrect assignement of space group: Now, some questions arise: 1) does the pseudo translation exist or do not? 2) if translational NCS is present, could it mimic a P41/P42/P43 space group? 3) in any case, any advices in order to properly deal with tNCS will be of great help? I thank you in advance Best wishes -- Fulvio Saccoccia, PhD Sapienza University of Rome Dept. of Biochemical Sciences "A. Rossi Fanelli" Piazzale Aldo Moro, 5 00185 Rome (Italy)

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rosetta installation problems
by Mengbin Chen 04 Jun '13

04 Jun '13

Dear Phenix Users, I am interested in using Rosetta to do MR, but unfortunately I have encountered some problems in installing it: I was able to get to the step where "scons: done building targets." showed up, and set up the environment by typing setenv /Users/yanghai/Desktop/rosetta-3.5 into the shell. I also filled out the "path to rosetta" in "Preferences" under "PHENIX GUI", which I believe it is correct. However, when I was ready to run MR-rosetta, it kept saying "cannot locate Rosetta programs. You may set the path in PHENIX GUI under preferences->Automation, or define the environment variable PHENIX_ROSETTA_PATH (recommended)". Any help would be apprecited! Thank you in advance, Mengbin Chen -- Mengbin Chen Department of Chemistry University of Pennsylvania

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